A h9 subtype avian influenza virus inactivated vaccine containing chicken α-interferon protein

A bird flu virus and interferon technology, applied in the direction of antiviral agents, peptide/protein components, medical preparations containing active ingredients, etc., can solve the problems such as no reports of joint preparation of vaccines, so as to improve the uniformity and increase the antibody Good level and safety

Active Publication Date: 2018-08-14
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] According to the existing research data, there is no report on the joint preparation of chicken α-interferon protein and H9 subtype avian influenza virus antigen
In 2004, Jia Lijun and others tested the immunoenhancing effect of expressed chicken gamma interferon on H5 subtype avian influenza vaccine. As a result, the titer of chicken gamma interferon combined immunization group was lower than that of inactivated vaccine alone

Method used

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  • A h9 subtype avian influenza virus inactivated vaccine containing chicken α-interferon protein
  • A h9 subtype avian influenza virus inactivated vaccine containing chicken α-interferon protein

Examples

Experimental program
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Effect test

Embodiment 1

[0013] Embodiment 1, the screening of antigenic strain (H9 subtype avian influenza virus QDY strain) for making seedlings

[0014] In 2013, the heart, spleen and other viscera, throat and cloaca swabs were aseptically collected from the affected chicken flock in Chengyang, Shandong, and were ground in a sterilized plate under aseptic conditions, and added in a ratio of 1:5 Physiological saline containing 4000 units / ml of double antibody was repeatedly frozen and thawed at -20°C for 3 times. Put the cotton swab directly into the physiological saline containing 10,000 units of double antibody, act for 4 hours at 2-8°C, centrifuge the viscera and cotton swab at 3500 rpm for 10 minutes, take the supernatant, mix and inoculate 10-day-old SPF chicken embryo 5 Pieces, 0.2ml / piece. Chicken embryos were incubated at 37°C, inspected once a day in the morning and evening, and the dead embryos were taken out in time and stored in a refrigerator at 4°C, and the dead embryos within 24 hour...

Embodiment 2

[0018] Embodiment 2: Preparation of antigen for seedling production

[0019] 1. Preparation of virus liquid for seedling production Inoculate 10-11-day-old SPF chicken embryos with virus seeds (QDY strain) for production through the allantoic cavity, 0.2ml per embryo, incubate at 36°C-37°C, and irradiate the embryos twice a day, Take the chicken embryos that died after 24 hours, harvest them at 96 hours regardless of whether they are dead, cool them at 4-8°C for 12-24 hours, harvest the chicken embryo liquid, mix them in a sterile container, and store them at 2-8°C.

[0020] 2. Preparation of chicken α-interferon protein for seedling production. Fermentation tank aeration culture, prepare medium according to 70% of the volume of the fermentation tank, adjust the pH value to 7.0-7.2 with 1mol / L NaOH, add 0.02% of the total volume of the medium foaming agent. After sterilization, inoculate the strains for production with 1% to 2% inoculation amount, add 10% kanamycin sulfate in...

Embodiment 3

[0028] One, the preparation of inactivated vaccine: the semi-finished product antigen after passing the inspection is carried out vaccine preparation (in the following preparation, each liquid component is calculated by volume ratio).

[0029] (1) Oil phase preparation Take 95 parts of veterinary white oil and 1 part of aluminum stearate, put them in the oil phase preparation tank and heat to 80°C, then add 805 parts of Siben-805, and when the temperature reaches 115°C, keep 30min, cool down and set aside.

[0030] (2) Preparation of aqueous phase Dilute chicken α-interferon protein with normal saline to 2×10 4.0 ~2×10 6.0 Unit / 0.1ml; Dilute the inactivated H9 subtype avian influenza virus solution with normal saline to not less than 2×10 6.0 EID 50 / 0.1ml. Take chicken α-interferon protein liquid and inactivated H9 subtype avian influenza virus liquid respectively, and mix them to prepare antigenic liquid for vaccine preparation. Take 5 parts of sterilized Tween-80 and a...

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Abstract

The invention provides an H9 subtype avian influenza virus inactivated vaccine including chicken a-interferon protein. The H9 subtype avian influenza virus inactivated vaccine comprises an antigen and a vaccine adjuvant, and the adopted antigen is an inactivated H9 subtype avian influenza virus QDY1 strain including the chicken a-interferon protein. A screened H9 subtype avian influenza virus has the advantages of being small in virulence and high in immunogenicity. The H9 subtype avian influenza virus (QDY strain) is inoculated into a chick embryo, then collects virus liquid, and is mixed with the extracted and purified chicken a-interferon protein after inactivation of formaldehyde solutions, and an oil adjuvant is added for emulsification to manufacture the vaccine. The prepared vaccine can stimulate an organism to generate an antibody fast, the level of the antibody is improved, the persistent period of the antibody is prolonged, and diseases caused by the H9 subtype avian influenza virus is prevented. The vaccine has the advantages of being efficient and good in safety.

Description

technical field [0001] The invention belongs to the technical field of biological veterinary medicine, in particular to a novel H9 subtype avian influenza vaccine containing chicken α-interferon protein. Background technique [0002] H9 subtype avian influenza is a low-pathogenic disease, but mixed infection with other pathogens, especially Escherichia coli, can increase its pathogenicity, cause a serious decline in egg production and a significant increase in mortality, and bring serious harm to the poultry industry. The existing avian influenza (subtype H9) vaccine has been widely used clinically, but there are problems of low antibody level and antibody uniformity, which affect the prevention and control effect of the vaccine. [0003] Interferon is a kind of low-molecular-weight soluble multifunctional glycoprotein produced by T lymphocytes and NK cells, which can enhance the function of cytotoxic T cells, promote the differentiation and activation of B cells, and induce...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/145A61K39/39A61P31/16A61K38/21
Inventor 刘新文杜元钊郭伟伟胡潇王秀丽宫晓
Owner YEBIO BIOENG OF QINGDAO
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